Biosensors and Nanotheranostics
Bionanotechnology, Drug Delivery, Therapeutics | online ISSN 3064-7789
159
Citations
101.4k
Views
64
Articles
Volume 4 Number 1 2025
REVIEWS (Open Access)
Electrochemical Biosensors for Single-Cell Microfluidics: Trapping and Detection Strategies and the Reporting Gap Limiting Clinical Translation
Lamiah Hossain 1*
Biosensors and Nanotheranostics 4 (1) 1-10 https://doi.org/10.25163/biosensors.4110710
Submitted: 10 May 2025 Revised: 05 July 2025 Accepted: 12 July 2025 Published: 14 July 2025
Abstract
Single-cell analysis is now central to modern diagnostics because bulk measurements average away the cellular heterogeneity that often drives disease. Electrochemical biosensors integrated with microfluidic architectures that manipulate femtoliter volumes provide one of the most direct routes to capturing this heterogeneity at single-cell resolution. This review synthesizes the literature on microfluidic electrochemical biosensing platforms for single-cell analysis, focusing on cell-trapping mechanisms, biorecognition immobilization strategies, and detection modalities. Rather than pooling quantitative outcomes across studies, we compare the qualitative strengths, trade-offs, and translational barriers reported across hydrodynamic, acoustic, dielectrophoretic, and optical trapping approaches, and across amperometric, fluorescence-based, and mass-spectrometric detection schemes. Hydrodynamic and dielectrophoretic trapping consistently achieve higher capture efficiency at the cost of throughput, while acoustic approaches are consistently reported as gentler on cell viability. Hybrid detection strategies that combine electrochemical readouts with fluorescence or mass spectrometry partially resolve the sensitivity-throughput trade-off, though inconsistent reporting standards across studies limit direct comparison. We argue that inconsistent reporting, not any single technical limitation, is the primary obstacle between proof-of-concept devices and routine clinical tools, particularly in liquid biopsy applications, and we identify where standardized reporting and cross-laboratory validation are most needed.
Keywords: Electrochemical biosensors; Microfluidics; Single-cell analysis; Cell trapping; Liquid biopsy
References
Aerts, J. T., Louis, K. R., Crandall, S. R., et al. (2014). Patch clamp electrophysiology and capillary electrophoresis-mass spectrometry metabolomics for single cell characterization. Analytical Chemistry, 86(6), 3203–3208. https://doi.org/10.1021/ac500168d
Antfolk, M., & Laurell, T. (2017). Continuous flow microfluidic separation and processing of rare cells and bioparticles found in blood. Analytica Chimica Acta, 965, 9–35. https://doi.org/10.1016/j.aca.2017.02.017
Arreguin-Campos, R., Jiménez-Monroy, K. L., Diliën, H., Cleij, T. J., van Grinsven, B., & Eersels, K. (2021). Imprinted polymers as synthetic receptors in sensors for food safety. Biosensors, 11(2), 46. https://doi.org/10.3390/bios11020046
Ban, E., Chae, D. K., & Song, E. J. (2013). Determination of micro-RNA in cardiomyoblast cells using CE with LIF detection. Electrophoresis, 34(4), 598–604. https://doi.org/10.1002/elps.201200442
Bartlett, P. N. (2008). Bioelectrochemistry: Fundamentals, experimental techniques and applications. John Wiley & Sons. https://doi.org/10.1002/9780470753842
Bhattacharya, S., Chao, T. C., Ariyasinghe, N., et al. (2014). Selective trapping of single mammalian breast cancer cells by insulator-based dielectrophoresis. Analytical and Bioanalytical Chemistry, 406(7), 1855–1865. https://doi.org/10.1007/s00216-013-7598-2
Bigi, A., Cojazzi, G., Panzavolta, S., Rubini, K., & Roveri, N. (2001). Mechanical and thermal properties of gelatin films at different degrees of glutaraldehyde crosslinking. Biomaterials, 22, 763–768. https://doi.org/10.1016/S0142-9612(00)00236-2
Brady, D., & Jordaan, J. (2009). Advances in enzyme immobilization. Biotechnology Letters, 31, 1639–1650. https://doi.org/10.1007/s10529-009-0076-4
Broyles, C. N., Robinson, P., & Daniels, M. J. (2018). Fluorescent, bioluminescent, and optogenetic approaches to study excitable physiology in the single cardiomyocyte. Cells, 7(6), 51. https://doi.org/10.3390/cells7060051
Burrow, G. M., & Gaylord, T. K. (2011). Multi-beam interference advances and applications: Nano-electronics, photonic crystals, metamaterials, subwavelength structures, optical trapping, and biomedical structures. Micromachines, 2(2), 221–257. https://doi.org/10.3390/mi2020221
Chen, A., Byvank, T., Chang, W. J., et al. (2013). On-chip magnetic separation and encapsulation of cells in droplets. Lab on a Chip, 13(6), 1172–1181. https://doi.org/10.1039/c2lc41201b
Chen, Y., Li, S., Gu, Y., et al. (2014). Continuous enrichment of low-abundance cell samples using standing surface acoustic waves (SSAW). Lab on a Chip, 14(5), 924–930. https://doi.org/10.1039/C3LC51001H
De Wael, K., De Belder, S., Van Vlierberghe, S., Van Steenberge, G., Dubruel, P., & Adriaens, A. (2010). Electrochemical study of gelatin as a matrix for the immobilization of horse heart cytochrome c. Talanta, 82, 1980–1985. https://doi.org/10.1016/j.talanta.2010.08.019
De Wael, K., De Belder, S., Pilehvar, S., Van Steenberge, G., Herrebout, W., & Heering, H. A. (2012). Enzyme-gelatin electrochemical biosensors: Scaling down. Biosensors, 2(1), 101–113. https://doi.org/10.3390/bios2010101
Fang, Y. L., Wang, C. H., Chen, Y. S., et al. (2021). An integrated microfluidic system for early detection of sepsis-inducing bacteria. Lab on a Chip, 21(1), 113–121. https://doi.org/10.1039/D0LC00966K
García-Galán, C., Berenguer-Murcia, A., Fernández-Lafuente, R., & Rodrigues, R. C. (2011). Potential of different enzyme immobilization strategies to improve enzyme performance. Advanced Synthesis & Catalysis, 353, 2885–2904. https://doi.org/10.1002/adsc.201100534
Gil, E. D., & de Melo, G. R. (2010). Electrochemical biosensors in pharmaceutical analysis. Brazilian Journal of Pharmaceutical Sciences, 46(3), 375–391. https://doi.org/10.1590/S1984-82502010000300002
Gou, Y., Jia, Y., Wang, P., & Sun, C. (2018). Progress of inertial microfluidics in principle and application. Sensors, 18(6), 1762. https://doi.org/10.3390/s18061762
Huang, C. J., Liu, C. X., & Stakenborg, J. L. T. (2014). Single cell viability observation in cell dielectrophoretic trapping on a microchip. Applied Physics Letters, 104(1), 013703. https://doi.org/10.1063/1.4861135
Junker, J. P., & van Oudenaarden, A. (2014). Every cell is special: Genome-wide studies add a new dimension to single-cell biology. Cell, 157(1), 8–11. https://doi.org/10.1016/j.cell.2014.02.010
Kalyan, S., Torabi, C., Khoo, H., Sung, H. W., Choi, S.-E., Wang, W., Treutler, B., Kim, D., & Hur, S. C. (2021). Inertial microfluidics enabling clinical research. Micromachines, 12(3), 257. https://doi.org/10.3390/mi12030257
Keithley, R. B., Rosenthal, A. S., Essaka, D. C., et al. (2013). Capillary electrophoresis with three-color fluorescence detection for the analysis of glycosphingolipid metabolism. Analyst, 138(1), 164–170. https://doi.org/10.1039/C2AN36286D
Lai, H. C., Wang, C. H., Liou, T. M., & Lee, G. B. (2014). Influenza A virus-specific aptamers screened by using an integrated microfluidic system. Lab on a Chip, 14(12), 2002–2013. https://doi.org/10.1039/C4LC00187G
Lao, Z., Xia, N., Wang, S., Xu, T., Wu, X., & Zhang, L. (2021). Tethered and untethered 3D microactuators fabricated by two-photon polymerization: A review. Micromachines, 12(4), 465. https://doi.org/10.3390/mi12040465
Larsen, S. T., Heien, M. L., & Taboryski, R. (2012). Amperometric noise at thin film band electrodes. Analytical Chemistry, 84(18), 7744–7749. https://doi.org/10.1021/ac301136x
Lin, L. Y., Chu, Y. S., Thiery, J. P., Lim, C. T., & Rodriguez, I. (2013). Microfluidic cell trap array for controlled positioning of single cells on adhesive micropatterns. Lab on a Chip, 13(4), 714–721. https://doi.org/10.1039/c2lc41070b
Liu, J., Yin, L. F., Dai, Y. R., Jiang, F., & Niu, J. F. (2012). Application of electrochemical enzyme biosensor in environmental pollution monitoring. Progress in Chemistry, 24(1), 131–143.
Liu, Y., & Wang, W. (2022). Multi-modal microfluidics (M3) for sample preparation of liquid biopsy: Bridging the gap between proof-of-concept demonstrations and practical applications. Micromachines, 13(2), 209. https://doi.org/10.3390/mi13020209
Mellors, J. S., Jorabchi, K., Smith, L. M., & Ramsey, J. M. (2010). Integrated microfluidic device for automated single cell analysis using electrophoretic separation and electrospray ionization mass spectrometry. Analytical Chemistry, 82(3), 967–973. https://doi.org/10.1021/ac902218y
Monosik, R., Ukropcova, D., Stredansky, M., & Sturdik, E. (2012). Multienzymatic amperometric biosensor based on gold and nanocomposite planar electrodes for glycerol determination in wine. Analytical Biochemistry, 421(1), 256–261. https://doi.org/10.1016/j.ab.2011.10.020
Neoh, K. H., Hassan, A. A., Chen, A., et al. (2018). Rethinking liquid biopsy: Microfluidic assays for mobile tumor cells in human body fluids. Biomaterials, 150, 112–124. https://doi.org/10.1016/j.biomaterials.2017.10.006
Qiu, J., Xu, J., Zhang, K., et al. (2020). Refining cancer management using integrated liquid biopsy. Theranostics, 10(5), 2374–2384. https://doi.org/10.7150/thno.40677
Ramser, K., & Hanstorp, D. (2010). Optical manipulation for single-cell studies. Journal of Biophotonics, 3(3), 187–206. https://doi.org/10.1002/jbio.200910050
Ren, K. N., Zhou, J. H., & Wu, H. K. (2013). Materials for microfluidic chip fabrication. Accounts of Chemical Research, 46(11), 2396–2406. https://doi.org/10.1021/ar300314s
Schubert, D. W., & Dunkel, T. (2003). Spin coating from a molecular point of view: Its concentration regimes, influence of molar mass and distribution. Materials Research Innovations, 7(5), 314–321. https://doi.org/10.1007/s10019-003-0270-2
Sierra, J., Marrugo-Ramírez, J., Rodríguez-Trujillo, R., et al. (2020). Sensor-integrated microfluidic approaches for liquid biopsy applications in early detection of cancer. Sensors, 20(5), 1317. https://doi.org/10.3390/s20051317
Sochol, R. D., Dueck, M. E., Li, S., Lee, L. P., & Lin, L. W. (2012). Hydrodynamic resettability for a microfluidic particulate-based arraying system. Lab on a Chip, 12(23), 5051–5056. https://doi.org/10.1039/c2lc40704c
Recommended articles
Advancing Personalized Cancer Care: Integrating CRISPR/Cas9 with Next-Generation Sequencing Technologies
Profiling of Autoantibodies as Biomarkers in Glioblastoma Multiforme (GBM) Patients
2
Save
Save
0
Citation
Citation
845
View
View
0
Share
Share